Fliess A, Wolfes H, Rosenthal A, Schwellnus K, Blöcker H, Frank R, Pingoud A
Nucleic Acids Res. 1986 Apr 25;14(8):3463-74. doi: 10.1093/nar/14.8.3463.
We have synthesized a series of oligonucleotides containing the EcoRI (GAATTC) or EcoRV (GATATC) recognition site within which or adjacent to which thymidine was substituted by uridine or derivatives of uridine. The effects of these substitutions on the rate of the EcoRI and EcoRV catalyzed cleavage reaction were investigated. Our results show that most of the substitutions within the site are quite well tolerated by EcoRI, not, however, by EcoRV. We conclude that the thymin residues most likely are not directly involved in the recognition process of the EcoRI reaction. In contrast, they are major points of contact, between substrate and enzyme in the EcoRV reaction. The effects of substitutions in the position adjacent to the recognition site is also markedly different for EcoRI and EcoRV. Here, EcoRI seems to be considerably more selective than EcoRV.
我们合成了一系列含有EcoRI（GAATTC）或EcoRV（GATATC）识别位点的寡核苷酸，在该识别位点内或其相邻位置，胸腺嘧啶被尿嘧啶或尿嘧啶衍生物取代。研究了这些取代对EcoRI和EcoRV催化的切割反应速率的影响。我们的结果表明，该位点内的大多数取代对EcoRI具有相当好的耐受性，但对EcoRV则不然。我们得出结论，胸腺嘧啶残基很可能不直接参与EcoRI反应的识别过程。相比之下，它们是EcoRV反应中底物与酶之间的主要接触点。识别位点相邻位置的取代对EcoRI和EcoRV的影响也明显不同。在这里，EcoRI似乎比EcoRV选择性更强。
